Use of carbon dioxide supplying means for muscle strengthening and method of increasing cattle meat thereby

ABSTRACT

Use of a carbon dioxide-supplying means for muscle strengthening makes it possible conveniently to strengthen a target muscle within a short period of time merely by allowing the target site to absorb carbon dioxide without loading any mechanical burden on the target muscle. By loading a mechanical burden on the target muscle, the muscle strengthening effect can be further enhanced and an additional effect of promoting the recovery from muscle fatigue can be achieved owing to the mechanical burden. By using the carbon dioxide-supplying means for muscle strengthening as described above, it is also possible to increase cattle meat.

CROSS REFERENCE

The present application is a 37 C.F.R. §1.53(b) divisional of, andclaims priority to, U.S. application Ser. No. 12/739,718, filed Apr. 23,2010. application Ser. No. 12/739,718 is the national phase under 35U.S.C. §371 of International Application No. PCT/JP2008/069351, filed onOct. 24, 2008. Priority is also claimed to Japanese Application No.2007-277524 filed on Oct. 25, 2007. The entire contents of each of theseapplications is hereby incorporated by reference.

TECHNICAL FIELD

The present invention relates to use of a carbon dioxide supplying meansfor muscle strengthening. More particularly, it relates to a musclestrengthening agent and a muscle strength decrease inhibitor containingcarbon dioxide as an active ingredient, a muscle training method usingthe carbon dioxide supplying means, and a method of increasing cattlemeat by using the carbon dioxide supplying means.

BACKGROUND ART

Patent Document 1 discloses that carbon dioxide is generally effectivefor the following symptoms (1) to (10) (see Patent Document 1):

(1) itching accompanying mucocutaneous diseases or mucocutaneousdisorders such as athlete's foot, insect bite, atopic dermatitis,nummular eczema, xeroderma, seborrheic eczema, urticaria, prurigo,housewives' eczema, acne vulgaris, impetigo, folliculitis, carbuncle,furunculosis, phlegmon, pyoderma, psoriasis, ichthyosis, palmoplantarkeratoderma, lichen, pityriasis, wound, burn, rhagades, erosion, andchilblain; mucocutaneous injuries such as decubitus ulcer, wound, burn,angular stomatitis, stomatitis, skin ulcer, rhagades, erosion,chilblain, and gangrene;(2) incomplete engraftment of skin grafts, skin flaps etc.;(3) dental diseases such as gingivitis, alveolar pyorrhea, dentureulcers, nigricans gingiva, and stomatitis;(4) skin ulcers, cryesthesia and numbness caused by peripheralcirculatory disorders such as thromboangitis obliterans,arteriolosclerosis obliterans, diabetic peripheral circulatorydisorders, and lower limb varicosis;(5) musculoskeletal diseases such as chronic rheumatoid arthritis,cervico-omo-brachial syndrome, myalgia, arthralgia and lumbago;(6) nervous system diseases such as neuralgia, polyneuritis, andsubacute myelo-optic neuropathy;(7) keratoses such as psoriasis, corns, calluses, ichthyosis,palmoplantar keratoderma, lichen, and pityriasis;(8) suppurative skin diseases such as acne vulgaris, impetigo,folliculitis, carbuncles, furuncles, phlegmon, pyoderma, and suppurativeeczema;(9) suppression of hair regrowth after depilation (treatment of unwantedhair); and(10) cosmetic troubles with the skin or hair such as freckles, roughskin, loss of clarity of the skin, loss of elasticity or luster of theskin, and loss of glossiness of the hair, and partial obesity.

In order to achieve an improvement of the above-mentioned symptoms, as acarbon dioxide supplying means, a composition for preparing a carbondioxide agent for external use (see Patent Documents 2 to 3), a carbondioxide composition for external use (see Patent Document 4), acomposition for preparing a carbon dioxide gel for external use (seePatent Document 5), a material for preparing a carbon dioxide agent forexternal use (see Patent Document 6), and a carbon dioxide externaladministration device (see Patent Documents 7 to 8) are disclosed.

Furthermore, Patent Document 9 discloses reduction of pain involved inmusculoskeletal disorders by transdermal introduction of carbon dioxide.

However, use of a carbon dioxide supplying means for musclestrengthening, and more particularly, a muscle strengthening agent and amuscle strength decrease inhibitor containing carbon dioxide as anactive ingredient, a muscle training method and a method of increasingcattle meat which use the carbon dioxide supplying means have not beenknown at all.

As a muscle strengthening method, muscle training in which a targetmuscle is contracted by the strength of 60% or more of the maximummuscle strength is common. In addition, kaatsu (pressure) muscletraining is proposed, which is said to be capable of enhancing musclestrength within a shorter period of time by pressurizing a target muscleso as to inhibit the blood flow during the exercise.

The muscle strengthening method by the above-mentioned muscle traininguses a phenomenon when muscle fatigue of a certain level or more isgenerated, the muscle strength is increased after a certain period oftime has passed. In this case, recovery from fatigue is necessarybecause if the recovery from muscle fatigue is slow, fatigueaccumulates, thus not only reducing the effect of the muscle trainingbut also tending to cause injury and the like. Therefore, in order toincrease the efficiency of muscle strengthening, the recovery fromfatigue is carried out by means of massage, lowfrequency therapyapparatus, warm bath, and the like, after the muscle training. However,it is reported that massage does not have an effect of promoting musclestrengthening (see Non-patent Document 1). Therefore, massage is notthought useful for the recovery from fatigue.

As a method of increasing the amount of the muscle which does not needmuscle training, a method of increasing the amount of the muscle byexternally applying a heat load to a living body has been proposed (seePatent Document 10). In the present invention, “muscle strengthening”and “increasing the amount of the muscle” are intended to have the samemeaning.

However, the method of increasing the amount of the muscle by applying aheat load described in Patent Document 10 requires accurate temperaturemanagement because the effective temperature for obtaining the effect ofincreasing the amount of the muscle (38 to 41° C.) approximates to thetemperature at which tissue cells are killed (43° C.). When a livingbody is warmed by the use of an external heat source, the temperature ofthe skin is the highest and the temperature becomes lower toward theinside of the living body. Therefore, it is not easy to heat only themuscle at the effective temperature for a certain period of time ormore.

Furthermore, the increasing of the amount of the muscle by this methodneeds such a long period of time as 14 weeks in order to increase, forexample, the biceps brachii by 4% even when muscle training is carriedout concurrently. Thus, this method is extremely ineffective.

Patent Document 1: Patent Document: Japanese Patent UnexaminedPublication No. 2000-319187 Patent Document 2: Patent Document: NationalPublication of International Patent Application No. 2002/80941 PatentDocument 3: Patent Document: National Publication of InternationalPatent Application No. 2006/80398 Patent Document 4: Patent Document:National Publication of International Patent Application No. 2003/57228Patent Document 5: Patent Document: National Publication ofInternational Patent Application No. 2005/16290 Patent Document 6:Patent Document: National Publication of International PatentApplication No. 2004/4745 Patent Document 7: Patent Document: NationalPublication of International Patent Application No. 2004/2393 PatentDocument 8: Patent Document: National Publication of InternationalPatent Application No. 2008/047829

Patent Document 9: Patent Document: U.S. Pat. No. 6,652,479B2

Patent Document 10: Patent Document: Japanese Patent UnexaminedPublication No. 2006-61687

Non-patent Document 1: Non-patent Document 1: Sven Joenhagen et al.,Sports Massage After Eccentric Exercise. The American Journal of SportsMedicine. 2004; 6: 1499-1503

DISCLOSURE OF THE INVENTION Problems to be Solved by the Invention

The present invention has an object to provide use of a carbon dioxidesupplying means for muscle strengthening, which makes it possible toobtain an effect within a short period of time in an easy and simplemanner. Specifically, the present invention has an object to provide amuscle strengthening agent and a muscle strength decrease inhibitorcontaining carbon dioxide as an active ingredient, as well as a muscletraining method and a method of increasing cattle meat which use thecarbon dioxide supplying means. The present invention has a furtherobject to concurrently provide an effect of promoting musclestrengthening and an effect of promoting recovery from fatigue in muscletraining, and the like.

Means for Solving the Problems

The present inventors have found that by allowing a living body on atarget site to absorb carbon dioxide with the use of a carbon dioxidesupplying means for muscle strengthening, the muscle strength isenhanced or muscle strength decrease is suppressed. The inventors havealso found that a carbon dioxide supplying means containing carbondioxide as an active ingredient is useful as a muscle strengtheningagent and a muscle strength decrease inhibitor. In addition, theinventors have found a muscle training method and a method of increasingcattle meat which use the carbon dioxide supplying means. Thus, theinventors have completed the present invention.

That is to say, the present invention provides use of a carbon dioxidesupplying means for muscle strengthening.

According to the present invention, it is possible to promote musclestrengthening and recovery from fatigue in a target site concurrently.

In the present invention, the muscle strengthening agent containingcarbon dioxide as an active ingredient includes any carbondioxide-containing agents for external use for muscle strengthening.Examples of the muscle strengthening agent include gaseous carbondioxide, carbon dioxide contained in a viscous composition, carbondioxide dissolved in water of water vapor, carbon dioxide dissolved in asolvent capable of dissolving carbon dioxide, and the like.Administration techniques thereof may include nebulization, application,patch, and the like. The administration techniques are not particularlylimited as long as they are methods by which carbon dioxide can besupplied to a target site. Furthermore, the muscle strength decreaseinhibitor containing carbon dioxide as an active ingredient includes anycarbon dioxide-containing agents for external use for suppressing thedecrease in muscle strength. Examples of the muscle strength decreaseinhibitor include gaseous carbon dioxide, carbon dioxide contained in aviscous composition, carbon dioxide dissolved in water of water vapor,carbon dioxide dissolved in a solvent capable of dissolving carbondioxide, and the like. Administration techniques thereof may includenebulization, application, patch, and the like. The administrationtechniques are not particularly limited as long as they are methods bywhich carbon dioxide can be supplied to a target site.

The present invention is a muscle strengthening agent and a musclestrength decrease inhibitor containing carbon dioxide as an activeingredient.

The present invention is the muscle strengthening agent and the musclestrength decrease inhibitor by which carbon dioxide is locallyadministered by using one or more carbon dioxide supplying meansselected from the group consisting of a carbon dioxide externaladministration device, a carbon dioxide agent for external use, acarbonated spring, water vapor containing carbon dioxide, and anintra-tissue injection of carbon dioxide.

The present invention is the muscle strengthening agent and the musclestrength decrease inhibitor by which carbon dioxide is transdermallyadministered by using the carbon dioxide external administration device.

The present invention is the muscle strengthening agent and the musclestrength decrease inhibitor using the above mentioned carbon dioxideexternal administration device, which is characterized by including asealing enclosure member capable of sealing a body surface from theoutside air, a supplying device for supplying carbon dioxide into theinside of the sealing enclosure member, and an absorption aid forassisting transdermal or transmucosal absorption of the carbon dioxideinside the sealing enclosure member.

The present invention is a muscle training method including acombination of transdermal absorption of carbon dioxide using one ormore carbon dioxide supplying means selected from the group consistingof a carbon dioxide external administration device, a carbon dioxideagent for external use, a carbonated spring, water vapor containingcarbon dioxide, and an intra-tissue injection of carbon dioxide, andapplying a load to a target muscle and/or inhibiting the blood flow tothe muscle.

The present invention provides a muscle training method including acombination of a transdermal absorption of carbon dioxide using a carbondioxide external administration device characterized by including asealing enclosure member capable of sealing a body surface from theoutside air, a supplying device for supplying carbon dioxide into theinside of the sealing enclosure member, and an absorption aid forassisting transdermal or transmucosal absorption of the carbon dioxideinside the sealing enclosure member; and applying a load to a targetmuscle and/or inhibiting the blood flow to the muscle.

The present invention is a muscle training method including acombination of a transdermal absorption of carbon dioxide using a carbondioxide external administration device characterized by including asealing enclosure member capable of sealing a body surface of a human oran animal from the outside air, a supplying device for supplying carbondioxide into the inside of the sealing enclosure member, and aperspiration promoting means for promoting perspiration on the bodysurface inside the sealing enclosure member; and applying a load to atarget muscle and/or inhibiting the blood flow to the muscle.

The present invention is a method of increasing cattle meatcharacterized by using one or more carbon dioxide supplying meansselected from the group consisting of a carbon dioxide externaladministration device, a carbon dioxide agent for external use, acarbonated spring, water vapor containing carbon dioxide, and anintra-tissue injection of carbon dioxide.

Advantages of the Invention

A muscle strengthening agent and a muscle strength decrease inhibitor ofthe present invention make it possible to enhance the muscle strength orsuppress muscle strength decrease within a short period of time in aneasy and simple manner when they are applied to aged persons whosemuscle strength is weakened or patients who have pain in the lower back,the elbow or the knee, who have difficulty in performing muscletraining. The enhancement of the muscle strength or suppression ofmuscle strength decrease increases the physical ability for behavior,and therefore enables independence of behavior to be improved, the areaof movement to be enlarged, and the quality of life to be improved inaged persons or patients whose muscle strength is weakened. Since themuscle strengthening agent and the muscle strength decrease inhibitor ofthe present invention can be easily applied to bedridden patients, andthe like, they can promote recovery in such patients and preventdisorders such as disuse atrophy. The muscle strengthening agent and themuscle strength decrease inhibitor of the present invention can preventmuscle strength decrease and arthrogryposis occurring at the time when afraction site or the like is fixed by applying a plastic cast with afunction of a sealing enclosure member described in Patent Document 7(Japanese Patent Unexamined Publication No. 2004-517345). When themuscle strengthening agent and/or the muscle strength decrease inhibitorof the present invention is applied during rehabilitation after fixingis completed, recovery of the muscle strength is promoted and the numberof dates required for rehabilitation can be shortened. In addition, themuscle strengthening agent and the muscle strength decrease inhibitor ofthe present invention are effective when the muscle strength isdecreased at the time of numbness by cerebrovascular disorder, spinalcord damage, bed rest after surgical operation, and the like.

As the muscle strengthening agent and the muscle strength decreaseinhibitor of the present invention, a portable carbon dioxidetransdermal absorption means such as a composition for preparing acarbon dioxide agent for external use and a carbon dioxide externaladministration device can be used. Therefore, it is possible to enhancethe muscle strength and/or to suppress muscle strength decrease easilyeven if specific facilities such as a gymnasium and a rehabilitationroom are not available.

The muscle strengthening agent and the muscle strength decreaseinhibitor of the present invention are not affected by the gravity.Therefore, when, for example, astronauts use the muscle strengtheningagent and the muscle strength decrease inhibitor during a rest orsleeping time, the time required to do exercise for preventingdeterioration of physical functions can be shortened. Thus, they canspend more time in their tasks such as scientific experimentsefficiently.

The muscle strengthening agent and the muscle strength decreaseinhibitor of the present invention not only promote muscle strengtheningin a short period of time by muscle training but also promote recoveryfrom fatigue in athletes, thus enabling their physical abilities to beimproved efficiently and safely.

Needless to say, since the muscle strengthening agent and the musclestrength decrease inhibitor of the present invention can be applied toanimals, for example, running ability of racehorses can be improvedefficiently and safely. Furthermore, use of the carbon dioxide supplyingmeans for muscle strengthening of the present invention can alsoincrease meat of cattle such as cow, pig, and sheep safely andefficiently.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a graph showing the change of the muscle strength of thequadriceps femoris when the muscle strength in the pre-ex (before anexercise load is given. The “ex” denotes a stretching exercise onisometric torque of the quadriceps femoris. The same is truehereinafter.) is defined as 100%;

FIG. 2 is a graph showing the change of the muscle strength of thehamstrings when the muscle strength in the pre-ex is defined as 100%;

FIG. 3 is a graph showing the muscle strength of the quadriceps femorisand the muscle strength of the hamstrings seven days after an exerciseload is given, when the muscle strength in the pre-ex is defined as100%;

FIG. 4 is a graph showing the muscle strength after the three-monthsexperiment in a case where an exercise load is not given;

FIG. 5 is a graph showing the ratio of lower limb muscle wet weight tobody weight of rat after the three-months experiment in a case where anexercise load is not given;

FIG. 6 is a graph showing amounts of proteins extracted from the muscleof rat lower limb of the three-months experiment in a case where anexercise load is not given; and

FIG. 7 is a graph showing the grip strength on day 3 after kaatsutraining is carried out.

BEST MODE FOR CARRYING OUT THE INVENTION

Use of a carbon dioxide supplying means for muscle strengthening inaccordance with the present invention is not particularly limited aslong as it allows a living body to absorb carbon dioxide. However, alocal absorption means is preferred. An example of the carbon dioxidesupplying means for allowing a living body to absorb carbon dioxide mayinclude the followings.

(1) A composition for preparing carbon dioxide agents for external use,for example, carbon dioxide agents for external use obtained from thefollowing a) and b): a) a composition for preparing a carbon dioxideagent for external use characterized by comprising a substancegenerating an acid after being hydrolyzed, a carbonate, a thickener, andwater as essential components, and further comprising a gelating agentbeing gelated by calcium ion, and a water-insoluble or poorly-solublecalcium salt (National Publication of International Patent ApplicationNo. 2006/80398), or b) a composition for preparing a carbon dioxideagent for external use comprising a granular material containing awater-soluble acid, a thickener, and a water-soluble dispersant as theessential components in which the thickener is mixed with thewater-soluble acid and the water-soluble acid and the water-solubledispersant; and a viscous composition containing a carbonate, water, anda thickener as the essential components, which is to be mixed with thegranular material at use (National Publication of International PatentApplication No. 2002-80941).

(2) A carbon dioxide composition for external use, characterized in thatcarbon dioxide is dissolved in a non-bubble state in a viscous materialcomprising at least water and a thickener, or a carbon dioxidecomposition for external use characterized by comprising at least afermentation microbe, a metabolite of the fermentation microbe, athickener, water, and carbon dioxide (National Publication ofInternational Patent Application No. 2003/57228).

(3) A carbon dioxide agents for external use obtained from a compositionfor preparing a carbon dioxide gel for external use characterized bycomprising the following granular material (A) and viscous material (B)to be mixed with the granular material (A):

(A) a granular material comprising a weak acid and a calcium iontrapping agent as essential components; and

(B) a viscous material comprising calcium carbonate, a gelling agentgelled by calcium ions and water as essential components (see, NationalPublication of International Patent Application No. 2005/16290).

(4) A carbon dioxide external administration device, characterized bycomprising a sealing enclosure member capable of sealing a body surfacefrom the outside air, a supply means for supplying carbon dioxide intothe inside of the sealing enclosure member, and an absorption aid thatassists transdermal or transmucosal absorption of the carbon dioxideinside the sealing enclosure member (National Publication ofInternational Patent Application No. 2004/02393).

(5) A carbon dioxide external administration device, characterized bycomprising a sealing enclosure member capable of sealing a body surfaceof a human or an animal from the outside air, a supply means forsupplying carbon dioxide into the inside of the sealing enclosuremember, and a perspiration promoting means for promoting perspiration onthe body surface inside the sealing enclosure member (NationalPublication of International Patent Application No. 2008/047829).

In addition, a carbonated spring such as an artificial carbonated springand water vapor containing carbon dioxide can also be used. The watervapor containing carbon dioxide denotes, for example, natural vapor thatcontains carbon dioxide and is erupted from underground, and water vaporwhite smoke that contains carbon dioxide, which is generated when dryice is put in the air. The water vapor containing carbon dioxide is usedfor hemorrhoid treatment, and the like, in for example, East Europe.Furthermore, a method of injecting carbon dioxide directly into targettissue with the use of a syringe, and the like, can be employed.Needless to say, the present invention is not limited to these carbondioxide supplying means, and may include any means that allow a targetsite to absorb carbon dioxide in an amount capable of promoting musclestrengthening. Among them, the carbon dioxide external administrationdevices described in the above-mentioned (4) or (5) are preferred.

The above-mentioned carbon dioxide supplying means may also be appliedto muscle training. When the use of the carbon dioxide supplying meansfor muscle strengthening of the present invention is employed in muscletraining, the absorption of carbon dioxide may be carried out at anytime from before, during or after the muscle training. However, theabsorption of carbon dioxide is preferably after the muscle trainingbecause an effect of recovery from fatigue is strong. In the presentinvention, the muscle training includes so-called kaatsu training, thatis, training while the muscle is pressurized by various methods.

As a concentration of carbon dioxide to be absorbed and absorption timemay vary depending upon carbon dioxide supplying means to be used. Inthe case where a carbon dioxide external administration device is used,the carbon dioxide concentration is preferably 300 ppm or more, and morepreferably 1,000 ppm or more. The absorption time may be preferablyabout five minutes or more, and more preferably ten minutes or more.Since carbon dioxide has highly safety, its absorption for a long periodof time does not matter. However, an expected effect can be obtained inabout 30 minutes.

When a carbon dioxide agent for external use, for example, thecomposition for external use mentioned above (2) is used, theconcentration of carbon dioxide to be dissolved is preferably 300 ppm ormore, and particularly preferably in the range from 1,000 ppm to thesaturation concentration.

Absorption frequency of carbon dioxide is preferably once or more aweek, and more preferably once or more a day.

When use of the carbon dioxide supplying means for muscle strengtheningof the present invention is carried out concurrently with muscletraining, it is preferable that absorption of carbon dioxide is carriedout every time muscle training is carried out. However, expected effectcan substantially be obtained even when the absorption of carbon dioxideis skipped once in several times. It is preferable that the absorptionof carbon dioxide is carried out even in days when muscle training isnot carried out because muscle strengthening and recovery from fatigueare promoted.

EXAMPLES

Hereinafter, the present invention will be specifically described withreference to Examples. However, the present invention is not limited tothese Examples.

Example 1

Promotion of Muscle Strengthening by Stretching Exercise on IsometricTorque of Legs (a carbon dioxide external administration device was usedas a carbon dioxide supplying means for muscle strengthening)

[Experiment Method]

Eight subjects including two subjects who played sports three times ormore a week (hereinafter, referred to as “athlete”) and six subjects whoplayed sports two times or less a week (hereinafter, referred to as“non-athlete”) participated in the experiment. Each subject didwarming-up by pedaling a gym bicycle for ten minutes. Next, each subjectdid a stretching exercise on isometric torque of the quadriceps femoris300 times or more by using training machine MyoretRZ-450 (Kawasaki HeavyIndustries, Ltd.). Thus, an exercise load was applied to the quadricepsfemoris. Then, the muscle strength of the quadriceps femoris and themuscle strength of the hamstrings to which an exercise load was notapplied were measured by using MyoretRZ-450.

After the training was completed, the carbon dioxide absorption aid 4described in Example 5 in Patent Document 7 (Japanese Patent UnexaminedPublication No. 2004-517345) was applied to one determined leg(hereinafter, referred to as a “treated leg”). The treated leg wascovered with an 80 cm-long polypropylene bag type sealing enclosuremember equipped with a check valve. Then, the member was filled withcarbon dioxide from a carbon dioxide gas cylinder and stood still forten minutes. The carbon dioxide concentration was set to 80% (800,000ppm). No treatment was given to the other leg (hereinafter, referred toas a “non-treated leg”). The training was carried out only once, thistime.

For three days from the next day after the training, each subject didnot do a stretching exercise on isometric torque of the quadricepsfemoris. Each subject allowed the treated leg to absorb carbon dioxidefor ten minutes once a day as mentioned above by using the carbondioxide supplying means for muscle strengthening of the presentinvention (by using a carbon dioxide external administration device asthe carbon dioxide supplying means). Thereafter, the muscle strength ofthe quadriceps femoris and the muscle strength of the hamstrings weremeasured by using MyoretRZ-450.

On day 4 and later after the experiment was started, neither absorptionof carbon dioxide nor stretching exercise on isometric torque of thequadriceps femoris was carried out. On day 7 after the experiment wasstarted, the muscle strengths of the quadriceps femoris and thehamstrings were measured by using MyoretRZ-450.

[Experiment Results]

On the day of starting the experiment, about 25% decrease of the musclestrength was observed in both legs after the exercise load was applied(see post-ex in FIG. 1) when the muscle strength before an exercise loadwas applied (pre-ex) was defined as 100%.

During the absorption of carbon dioxide after the exercise load wasapplied, blood pressure and heart rate of the subjects did not change.After carbon dioxide was absorbed, when the muscle strength of thequadriceps femoris was measured by using MyoretRZ-450, recovery of themuscle strength was faster in the treated leg than in the non-treatedleg (see post-CO2 in FIG. 1).

On days 2 to 4 of the experiment, when the muscle strength of thequadriceps femoris was measured by using MyoretRZ-450, musclestrengthening was observed in both legs. The muscle strengthening waspromoted in the treated leg as compared with in the non-treated leg (seeon days 2, 3, and 4 in FIG. 1).

On day 7 of the experiment, when the muscle strength of the quadricepsfemoris was measured, only slight muscle strengthening was observed inthe non-treated leg. Meanwhile, in the treated leg, although theabsorption of carbon dioxide had been stopped, 20% muscle strengtheningwas observed. In particular, in the non-athletes, about 30% musclestrengthening was observed (see the result on day 7 in FIG. 1).

On the other hand, also in the hamstrings to which an exercise load wasnot given, about 15% muscle strengthening was observed in the treatedleg while muscle strengthening of the hamstrings was not observed in thenon-treated leg (see the result on day 7 in FIG. 2). As is apparent fromFIG. 3, the muscle strengthening was statistically significant at P=5%.Furthermore, as is apparent from FIG. 2, in particular, in the case ofnon-athletes, about 20% muscle strengthening was observed.

Example 2

Promotion of Muscle Strengthening with No Exercise Load Applied (acarbon dioxide external administration device was used as a carbondioxide supplying means for muscle strengthening)

[Experiment Method]

Thirteen subjects (all males, aged from 27 to 43) participated in theexperiment. In each subjects, the absorption aid 4 of carbon dioxidedescribed in Example 5 in Patent Document 7 (Japanese Patent UnexaminedPublication No. 2004-517345) was given to one determined leg(hereinafter, referred to as a “treated leg”). The treated leg wascovered with an 80 cm-long polypropylene bag type sealing enclosuremember equipped with a check valve. Then, carbon dioxide was filled inthe member from a carbon dioxide gas cylinder and stood still for tenminutes. The carbon dioxide concentration was set to 80% (800,000 ppm).The treatment was carried out at least once a week for three months. Notreatment was given to the other leg (hereinafter, referred to as a“non-treated leg”). Before and three months after the experiment wasstarted, the muscle strengths of the treated leg and the non-treated legwere measured (the muscle strengths of the quadriceps femoris and thehamstrings were measured by using MyoretRZ-450).

[Experiment Results]

On month 3, about 15% muscle strengthening was observed in thequadriceps femoris and the hamstrings in the treated leg as comparedwith the non-treated leg. The muscle strengthening were statisticallysignificant at P=1% in the quadriceps femoris and P=5% in the hamstrings(see FIG. 4).

Example 3

Increase in Amount of Rat Muscle with No Exercise Load Applied (a carbondioxide external administration device was used as a carbon dioxidesupplying means for muscle strengthening)

[Experiment Method]

Nine male Wister rats (5-week old at the time when the experiment wasstarted) were used. The rats were divided into two groups: a group ofrats that underwent absorption of carbon dioxide (hereinafter, referredto as a “CO2 group”) and a group of rats that did not undergo absorptionof carbon dioxide (hereinafter, referred to as a “CONTROL group”).

In the CO2 group, the hair was removed from the right and left lowerthighs of six rats under anesthesia. Then, the absorption aid 4 ofcarbon dioxide described in Example 5 in Patent Document 7 (JapanesePatent Unexamined Publication

No. 2004-517345) was applied to only the right lower thigh (hereinafter,referred to as a “treated limb”). Next, the rat lower body was coveredwith a polyethylene bag type sealing enclosure member provided with ahole through which the left hind limb comes out so that only the righthind limb absorbs carbon dioxide. Then, carbon dioxide was filled in thesealing enclosure member by using a carbon dioxide gas cylinder so as toinflate the sealing enclosure member. In order to always secure asufficient amount of carbon dioxide in the sealing enclosure member,carbon dioxide was added at any time, so that the sealing enclosuremember was filled with carbon dioxide for ten minutes. The carbondioxide concentration inside the sealing enclosure member was set to100%. The above-mentioned treatment was carried out twice a week forthree months. To the left lower thigh, only removing of hair was carriedout and other treatment was not carried out (hereinafter, referred to as“non-treated leg”).

In the CONTROL group, only anesthesia was carried out and absorption ofcarbon dioxide was not carried out.

Three months later, the rat body weights were measured and the musclewas collected from the right and left lower thighs. The muscle wetweight of the collected muscle was measured, followed by detectinggrowth a acceleration indicating proteins of muscle fibers, MyoD andMyogenin (see Atsuko HARA et al., “Expression of myosin heavy chainisoforms, myogenin and MyoD in the course of regeneration of skeletalmuscle,” Journal of Health Sciences of Hiroshima University, Vol. 2 (1):12-18, 2002), and IGF-IR [see Junichi SUZUKI et al., “Effect ofendurance training with L-arginine supplementation on expression ofIGF-1 receptor in rat skeletal muscles” Research for Winter Sports, (theProceedings of Research and Education Center for Winter Sports, HokkaidoUniversity of Education), Vol. 8 (1) 1-7, 2005], by Western blotting.The resultant bands were analyzed on the analysis software (NIH-Image)so as to quantify each growth acceleration indicating protein. Theresultant ratio of the muscle wet weight/body weight thus obtained isshown in FIG. 5 and the amount of each protein is shown in FIG. 6.

[Experiment Results]

As is apparent from FIG. 5, when the muscle wet weight and the bodyweight were measured, an increase in the muscle wet weight per bodyweight was observed in the CO2 group as compared with the CONTROL group.Furthermore, in the CO2 group, a remarkable increase in the muscle wetweight was observed in the treated leg as compared with the non-treatedleg.

As is apparent from FIG. 6, the amount of the growth accelerationindicating protein extracted from the muscle of the CO2 group wasincreased as compared with that of the CONTROL group.

These experiment results show that the muscle wet weight is increased bythe absorption of carbon dioxide, and the increase is brought about bythe growth acceleration of the muscle fiber.

Example 4

Promotion of Kaatsu Training Effect (a carbon dioxide externaladministration device was used as a carbon dioxide supplying means formuscle strengthening)

[Experiment Method]

Two subjects (one male and one female) participated in the experiment.The experiment was carried out according to Abe T. et al. (2006)Electromyographic responses of arm and chest muscle during bench pressexercise with and without KAATSU. Int. J. Kaatsu Training Res. 2:15-18.The grip strengths of both hands of each subject were measured beforethe experiment was started. Next, the absorption aid 4 of carbon dioxidedescribed in Example 5 in Patent Document 7 (Japanese Patent UnexaminedPublication No. 2004-517345) was applied to the entire left arm of eachsubject, and the left arm was covered with an 80 cm-long polypropylenebag type sealing enclosure member equipped with a check valve. A cuff ofblood pressure manometer was worn on the upper arm with the sealingenclosure member applied. Firstly, the upper arm was pressurized with acuff pressure of 50 mmHg for 30 seconds. Next, the pressure was releasedonce for 10 seconds, and then the upper arm was pressurized whilegradually increasing the cuff pressure to 70 mmHg. When the cuffpressure reached 70 mmHg, the pressure was fixed. Then, carbon dioxidewas filled in the sealing enclosure member from a carbon dioxide gascylinder so as to inflate the sealing enclosure member. Immediatelythereafter, the subjects carried out an exercise of opening and closingthe palm 20 times quickly, and then took a rest for 15 seconds. Thisexercise and rest were repeated three times (hereinafter, this arm isreferred to as a “treated arm”). In order to always secure a sufficientamount of carbon dioxide in the sealing enclosure member, carbon dioxidewas added at any time. Ten minutes after the initial pressurizing, thecuff pressure was released, and the sealing enclosure member wasremoved.

In the right arm, only pressurizing and exercise of opening and closingthe palm were carried out under the same condition as in the left arm,and absorption of carbon dioxide was not carried out (hereinafter, thisarm is referred to as a “non-treated arm”).

Three days after the experiment, the grip strength of both hands of eachsubject was measured.

[Experiment Results]

As is apparent from FIG. 7, it was shown that absorption of carbondioxide promoted the muscle strengthening effect by pressurizing. Ascompared with the case in which only pressurizing was carried out byusing a carbon dioxide supplying means for muscle strengthening, almosttwo-times muscle strengthening rate was shown. Training of onlypressurizing and an exercise of opening and closing the palm accompaniesa sever muscle pain or fatigue. However, in the left hand, which hadundergone the same training while allowing carbon dioxide to be absorbedwith the use of the carbon dioxide supplying means for musclestrengthening, the muscle pain and fatigue were reduced.

INDUSTRIAL APPLICABILITY

A muscle strengthening agent and a muscle strength decrease inhibitor ofthe present invention containing carbon dioxide as an active ingredientmake it possible to enhance the muscle strength or to suppress musclestrength decrease in the muscle in a target site in an easy and simplemanner within a short period of time merely by allowing a target muscleto absorb carbon dioxide without applying any mechanical loads to thetarget muscle. Furthermore, the muscle strengthening agent and themuscle strength decrease inhibitor of the present invention containingcarbon dioxide as an active ingredient are useful as a muscle trainingmethod in promoting muscle strengthening or suppressing the musclestrength decrease in a combination of applying a load to a target muscleand/or inhibiting the blood flow. Use of the carbon dioxide supplyingmeans for muscle strengthening of the present invention makes itpossible to increase an amount of cattle meat.

1. A muscle strengthening agent or a muscle strength decrease inhibitorcomprising carbon dioxide as an active ingredient, in which carbondioxide is transdermally administrated at a target site.
 2. The musclestrengthening agent or a muscle strength decrease inhibitor according toclaim 1, selected from the group of gaseous carbon dioxide, a carbondioxide agent for external use, a carbonated spring and water vaporcontaining carbon dioxide.
 3. The muscle strengthening agent or a musclestrength decrease inhibitor according to claim 1, selected from thegroup of gaseous carbon dioxide and water vapor containing carbondioxide.
 4. The muscle strengthening agent or a muscle strength decreaseinhibitor according to claim 3, wherein carbon dioxide is administratedwith a carbon dioxide external administration device characterized bycomprising a sealing enclosure member capable of sealing a body surfacefrom the outside air, a supply means for supplying carbon dioxide intothe inside of the sealing enclosure member, and an absorption aidcomprised of a viscous material containing a thickener and water whichassists transdermal or transmucosal absorption of the carbon dioxideinside the sealing enclosure member.
 5. The muscle strengthening agentor a muscle strength decrease inhibitor according to claim 4, whereinthe thickener is selected from sodium alginate and sodiumcarboxymethylcellulose.
 6. The muscle strengthening agent or a musclestrength decrease inhibitor according to claim 1, selected from thegroup of gaseous carbon dioxide and a carbon dioxide agent for externaluse.
 7. The muscle strengthening agent or a muscle strength decreaseinhibitor according to claim 6, wherein the concentration of carbondioxide is equal to or over 300 ppm.
 8. A kit comprising two or more ofan agent or an inhibitor selected from the muscle strengthening agent orthe muscle strength decrease inhibitor according to any of claims 1-7.